Abstract: Blends of enzymes of the same enzyme class (same EC number), such as beta-glucuronidase enzyme blends, are provided that exhibit synergistic levels of activity across a range of substrates as compared to the activity levels of each enzyme in the blend individually. The blends also may exhibit a greater effective substrate range, as well as greater effective pH and/or temperature ranges as compared to single enzyme preparations. Predictive methods for determining optimal enzyme blends, methods for preparing enzyme blends, enzyme blend compositions, enzyme blend formulations and methods of using such enzyme blends are provided.
Type:
Grant
Filed:
December 10, 2021
Date of Patent:
November 7, 2023
Assignee:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
Inventors:
John Tomashek, Caleb Reece Schlachter, Pongkwan Sitasuwan, Lim Andrew Lee
Abstract: Chimeric and other variant ?-glucuronidase enzymes with enhanced properties as compared to unmodified enzyme are provided. The enzymes of the invention advantageously exhibit enhanced enzymatic activity, enhanced substrate range, enhanced pH range, enhanced temperature range and/or enhanced enzyme stability. Methods of using the variant enzymes for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.
Type:
Grant
Filed:
October 8, 2019
Date of Patent:
August 23, 2022
Assignee:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
Inventors:
Caleb Reece Schlachter, John Tomashek, Lim Andrew Lee
Abstract: Blends of enzymes of the same enzyme class (same EC number), such as beta-glucuronidase enzyme blends, are provided that exhibit synergistic levels of activity across a range of substrates as compared to the activity levels of each enzyme in the blend individually. The blends also may exhibit a greater effective substrate range, as well as greater effective pH and/or temperature ranges as compared to single enzyme preparations. Predictive methods for determining optimal enzyme blends, methods for preparing enzyme blends, enzyme blend compositions, enzyme blend formulations and methods of using such enzyme blends are provided.
Type:
Grant
Filed:
July 31, 2019
Date of Patent:
March 8, 2022
Assignee:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
Inventors:
John Tomashek, Caleb Reece Schlachter, Pongkwan Sitasuwan, Lim Andrew Lee
Abstract: The present invention provides a disposable pipette tip for dispersive solid phase extraction (SPE) that allows for rapid, automatable purification of large biomolecules, such as nucleic acids, proteins and polypeptides without the need for additional tools such as centrifuges, magnetic plates or vacuum manifolds. The pipette tip is designed for optimal biomolecule isolation while maintaining sample integrity. Optimized methods of using the dispersive pipette extraction system for isolation of large biomolecules are also provided.
Type:
Grant
Filed:
May 25, 2017
Date of Patent:
November 2, 2021
Assignee:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
Inventors:
Brian Todd Mullis, Gary Horvath, Lim Andrew Lee, Sunil Hwang, William Edward Brewer
Abstract: Formulations of ?-glucuronidase enzymes that exhibit long-term stability at both high and low temperatures as either aqueous or lyophilized formulations are provided. The formulations of the invention comprise a ?-glucuronidase enzyme, such as a mutant ?-glucuronidase enzyme, an amphoteric compound, L-histidine,?-alanine and a sugar. Methods of preparing the formulations, as well as methods of using the formulations for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.
Type:
Application
Filed:
January 20, 2017
Publication date:
January 23, 2020
Applicant:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
Inventors:
Lim Andrew LEE, Pongkwan Nikki SITASUWAN, Margarita MARINOVA
Abstract: Mutated ?-glucuronidase enzymes with enhanced enzymatic activity and thermostability as compared to wild type enzyme are provided. The enzymes of the invention advantageously allow for accurate analysis of bodily samples for the presence of drugs in 30 minutes or less, as compared to the several hours needed using prior enzyme preparations. Methods of using the mutated enzymes for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.
Abstract: Mutated Lactobacillus brevis strain 269Y ?-glucuronidase enzymes with enhanced enzymatic activity at low pH (e.g., below pH 6.8), as well as enhanced thermostability as compared to wild type enzyme are provided. The enzymes of the invention advantageously allow for accurate analysis of bodily samples for the presence of drugs at low pH and in 30 minutes or less, as compared to the several hours needed using prior enzyme preparations. Methods of using the mutated enzymes for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.
Type:
Grant
Filed:
March 21, 2016
Date of Patent:
March 6, 2018
Assignee:
INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS
Inventors:
Jia Yang, Gary Horvath, Pongkwan Sitasuwan, Margarita Marinova, Qian Wang, Lim Andrew Lee
Abstract: Mutated Staphylococcus sp. RLH1 ?-glucuronidase enzymes with enhanced enzymatic activity and thermostability as compared to wild type enzyme are provided. The enzymes of the invention advantageously allow for accurate analysis of bodily samples for the presence of drugs in 30 minutes or less, as compared to the several hours needed using prior enzyme preparations. Methods of using the mutated enzymes for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.